P-selectin, also called GMP-140, CD62, or selectin P, is a 140kD adhesion molecule, expressed at the surface of activated cells, that mediates the interaction of activated endothelial cells or platelets with leukocytes. It is stored in secretory granules and expressed at the plasma membrane after cell activation. It is known to play an important role in atherosclerosis. The major ligand for P-selectin on leukocytes is P-selectin glycoprotein ligand-1 (PSGL-1). The standard product used in this kit is recombinant human P-Selectin, excluding intercellular P-Selectin and transmembrane domain. It has 730 amino acids sequence with the molecular mass of 80kD. Intended Use: P-Selectin BioAssay(TM) ELISA Kit, Human is a sandwich ELISA for quantitative detection of human P-Selectin in cell culture supernates, serum and plasma (heparin, EDTA, citrate). Range: 62.5-4000pg/ml Sensitivity: <5pg/ml Specificity: Natural and recombinant human P-Selectin Crossreactivity: No detectable crossreactivity with other relevant proteins Kit Components: 144043A: Microtiter Strips, 1x96 wells 144043B: P-Selectin Standard, 2x10ng 144043C: Anti-human P-Selectin (Biotin), 130ul USB Cat No Kit Component 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml Assay Summary: 1. Add samples and standards and incubate the plate at 37C for 90 minutes. Do not wash. 2. Add biotinylated antibodies and incubate the plate at 37C for 60 minutes. Wash plate 3 times with 0.01M TBS. 3. Add ABC working solution and incubate the plate at 37C for 30 minutes. Wash plate 5 times with 0.01M TBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 25-30 minutes. 5. Add TMB stop solution and read.. Storage and Stability: Store *144043B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
